Chemiluminescence resonance energy transfer imaging on magnetic particles for single-nucleotide polymorphism detection based on ligation chain reaction

被引:40
作者
Bi, Sai [1 ]
Zhang, Zhipeng [2 ]
Dong, Ying [2 ]
Wang, Zonghua [1 ]
机构
[1] Qingdao Univ, Collaborat Innovat Ctr Marine Biomass Fiber Mat &, Shandong Sino Japanese Ctr Collaborat Res Carbon, Growing Base State Key Lab,Lab Fiber Mat & Modern, Qingdao 266071, Peoples R China
[2] Qingdao Univ Sci & Technol, Key Lab Sensor Anal Tumor Marker, Minist Educ, Coll Chem & Mol Engn, Qingdao 266042, Peoples R China
基金
中国博士后科学基金; 中国国家自然科学基金;
关键词
Chemiluminescence resonance energy transfer; Ligation chain reaction; Magnetic particles; Single-nucleotide polymorphism; STRAND-DISPLACEMENT; HOMOGENEOUS ASSAY; GRAPHENE OXIDE; DNA; AMPLIFICATION; DNAZYME; CANCER; IMMUNOASSAY; BIOSENSORS; MUTATIONS;
D O I
10.1016/j.bios.2014.10.025
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
A novel ligation chain reaction (LCR) methodology for single-nucleotide polymorphism (SNP) detection was developed based on luminol-H2O2-horseradish peroxidase (HRP)-mimicking DNAzyme-fluorescein chemiluminescence resonance energy transfer (CRET) imaging on magnetic particles. For LCR, four unique target-complement probes (X and X*, YG and V) for the amplification of K-ras (G12C) were designed by modifying G-quadruplex sequence at 3'-end of YG and fluorescein at 5'-end of Y*. After the LCR, the resulting products of XYG/X*Y* with biotin-labeled X* were captured onto streptavidin-coated magnetic particles (SA-MPs) via specific biotin-SA interaction, which stimulated the CRET reaction from hemin/G-quadruplex-catalyzed luminol-H2O2 CL system to fluorescein. By collecting signals by a cooled low-light CCD, a CRET imaging method was proposed for visual detection and quantitative analysis of SNP. As low as 0.86 fM mutant DNA was detected by this assay, and positive mutation detection was achieved with a wild-type to mutant ratio of 10,000:1. This high sensitivity and specificity could be attributed to not only the exponential amplification and excellent discrimination of LCR but also the employment of SA-MPs. SA-MPs ensured the feasibility of the proposed strategy, which also simplified the operations through magnetic separation and separated the reaction and detection procedures to improve sensitivity. The proposed LCR-CRET imaging strategy extends the application of signal amplification techniques to SNP detection, providing a promising platform for effective and high-throughput genetic diagnosis. (C) 2014 Elsevier B.V. All rights reserved.
引用
收藏
页码:139 / 144
页数:6
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